大鼠聚腺苷二磷酸核糖聚合酶-1基因RNA干擾表達(dá)質(zhì)粒構(gòu)建與鑒定

《大鼠聚腺苷二磷酸核糖聚合酶-1基因RNA干擾表達(dá)質(zhì)粒構(gòu)建與鑒定》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在工程資料-天天文庫。

1、文章集號(hào)4095-2619(2012)01-0001-04大鼠聚腺昔二磷酸核糖聚合酶-1基因RNA干擾表達(dá)質(zhì)粒構(gòu)建與鑒定高羽辛心，黃明元2,劉林華?，梁海榮蔦唐煥文2摘要：目的構(gòu)建并篩選大貳聚腺藥二續(xù)咬換殊聚合基因的RNA干擾(RNAi)表達(dá)質(zhì)炷，為職業(yè)惶戎患的基因治療探索新途輕。方法根坍基因序列數(shù)據(jù)陣中報(bào)道.的PARP-I基因凈列及短發(fā)夬RNA(shRNA)設(shè)計(jì)原則，設(shè)計(jì)、合成用于構(gòu)建RWAi質(zhì)粒的眾核并酸，構(gòu)瑾4聲支姐PARPJRNAi用粒.醉切及測序鑒定正確后?以脂質(zhì)體U-pofecUminJYOOO介導(dǎo)轉(zhuǎn)染大風(fēng)骨牡間充質(zhì)干仙胞。48h后，采用逆轉(zhuǎn)敢聚合聲鏈反應(yīng)(RT?P(：R)技術(shù)檢

2、測輔染細(xì)胞中PARPJmRNA的轉(zhuǎn)錄水平，觴選有效的PARP-1R￥Ai質(zhì)粒。結(jié)果4種重組PARPJRNAi質(zhì)粒經(jīng)酹切及測序證明構(gòu)建正確"轉(zhuǎn)染后48h,轉(zhuǎn)染質(zhì)粒shRNA-PARP-1-532的細(xì)袍PARP-J基因柯制效果最明星，mRNA的轉(zhuǎn)錄水乎下隆了78%,可作為后裟實(shí)臉的有效質(zhì)杜:：＞結(jié)論成功構(gòu)建并捕il出PARP1基因的RNAi表達(dá)貞為探索PAKP-I基因在疾病中的作用莫芝了崔礎(chǔ)。關(guān)鎮(zhèn)詞：聚腺奪二輯絞根純聚合酶-】；隸發(fā)央RNA｛骨戕間充質(zhì)干佃胞；轉(zhuǎn)棗；大鼠中國分類號(hào)：R135文就標(biāo)識(shí)碼：AConstructionandscreefiingofRNAiexpressionvector

3、forratpoly(ADP-rib<>Ke)polymerase-!geneGAOYu-ting,HUANGMing-yuan,LIULin-hua,LIANGHai-n)ngtTANGHuan-wcn(DepartmentofToxicologyrSchoolofPublicHealth,JilinUniversity.Changchun130021,China)Abstract:ObjectiveTooonBtructandscreentheRNAiuxpn^onvec

4、eanovelrouteforthegenetherapyofoccupationaldiseases.MethodsTheoligonucleotideswmvdc?igi>rdtukdsynthesizedaccordingtothePARP-1genereportedinGenBankandthegenera]principleofshRNA(1疥ign.basedonwhich4m?combinantRNAiexpressionvcctoratargetingPARISIgenewereconstructedwidentifiedbyrestiictionanalysisandseq

5、uencing,thentransfectedtorathonemesenchymal^temcellsinmediationofUpofecuunine?2000.ThetranscriptionlevelsofPARP-1mKNAweredetenninedbywal?timercverectranscriptionpolymera^chainrcaclion(RT?P(：R)48hoursaftertrarusfretiontoscreenthevalidRNAiexpressionvectorstargetingPARPJgene.ResultsRestrictionanalysis

6、andsequencingprovedthat4recombinantRNAiexpressionvectorsUrgedry;PARP-1genewereconstructedcorrectly.ThetranscriptionlevelofPARP-1mRNAinbonemesenchymalstemcellstransfectedwithrecombinantplasmidsshRNA-PARP-1-532decreasedmoreremarkablr.hwasusedasthevalidRNAiexpressionvectoratargetingPARP-1geneforfurth

7、erstudy.ConclusionTheRNAiexpressionvectorforratPARP-Igenewassuccessfullyconstructedand乂reentd,whichlaidafoundiitionofstudyontheeffectofPARP-1glymeraj